CN107460133A - Dark color has every endogenetic fungus HS40 and its application in dendrobium candidum production - Google Patents
Dark color has every endogenetic fungus HS40 and its application in dendrobium candidum production Download PDFInfo
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- CN107460133A CN107460133A CN201710833508.3A CN201710833508A CN107460133A CN 107460133 A CN107460133 A CN 107460133A CN 201710833508 A CN201710833508 A CN 201710833508A CN 107460133 A CN107460133 A CN 107460133A
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
Abstract
The present invention relates to dark color to have every endogenetic fungus HS40 and its application in dendrobium candidum production.The invention discloses the dark color that deposit number is CGMCC NO.14345 to have every endogenetic fungus HS40, the endogenetic fungus can remarkably promote the growth and development of dendrobium candidum, it is inoculated with plate culture dendrobium candidum seedling plant height, stem diameter, plant fresh weight and the dry weight relatively control increase by 21.2%, 28.4%, 7.5% and 140.7% respectively of the bacterial strain, wherein, plant height and dry weight reach the pole level of signifiance with contrast difference；The relatively control increase by 400.0%, 94.8%, 56.8%, 15.8% respectively of dendrobium candidum Potted orchard tiller number, plant fresh weight, plant weights and the stem polyoses content of bacterial strain processing, wherein tiller number reaches the level of signifiance with contrast difference, plant fresh weight, dry weight and stem polyoses content then respectively reach the pole level of signifiance, illustrate that HS40 can effectively facilitate the growth of dendrobium candidum.
Description
【Technical field】
The present invention relates to biological technical field, more particularly to dark color has every endogenetic fungus HS40 and its produced in dendrobium candidum
On application.
【Background technology】
In recent years, world's microorganism strategy with the U.S. " national microorganism group plan " for representative is strengthened and has focused on agricultural
Great function of the microorganism in terms of solving the significant problems such as human health, environmental protection, the energy, agriculture microbe research is
As the strategy " highland " of international new round scientific and technological revolution.Meanwhile in recent years due to largely applying chemical fertilizer, agricultural chemicals to grain peace
Entirely, ecological environment and the caused harm of the utilization of resources have also caused national great attention, reduce chemical fertilizer, the applications of pesticide and raising
Chemical fertilizer utilization ratio turns into major issue urgently to be resolved hurrily in current China's agricultural production.Utilize the micro- life of plant symbiosis in nature
Thing promote crop growth meet country relevant policies, by as China's agriculture environmental protection, agricultural sustainable development one
Individual important directions.Dark color have every endogenetic fungus (DARK SEPTATE ENDOPHYTE, DSE) be plant symbiosis fungi typical case
One of represent, they can settle down in plant root but have separation to host's no pathogenicity, mycelia dark color, can be in root shape
Into architectural features such as " Microsclerotias ".Research shows that DSE can assign plant good development character, can carry with host's mutualistic symbiosis
The resistant to diseases and insects of high host and the resistance in stressful environmental, so as to reduce the use of chemical fertilizer, agricultural chemicals.This kind of fungi
With ecologicaI distribution is extensive, host range is wide and the characteristic of separable pure culture, this just determines them in agriculture sustainable development
It will be played an increasingly important role in exhibition, there is more wide application prospect.
Dendrobium candidum is a kind of rare rare medicinal and ornamental plant, is the rareness species in imminent danger that China is laid special stress on protecting.
In recent years, although dendrobium candidum tissue culture and artificial cultivation technique are rapidly progressed, the life of dendrobium candidum artificial cultivation
It is still the major obstacle for limiting dendrobium candidum industry development that long slow, fertilizer and pesticide, which is excessively applied,.At present, dendrobium candidum growth-promoting
The involved endogenetic fungus of research derives from dendrobium candidum in itself mostly, seldom from other host plants.The researchs such as Xie Ling are sent out
Existing, the endogenetic fungus 24L-4 bacterial strains for being isolated from camellia leaf portion of agricultural college of Japanese Ibaraki university have significantly to dendrobium candidum
Growth-promoting functions.Thus it is presumed that, China indigenous advantage plant equally contains may have good growth-promoting work(to dendrobium candidum
The endogenetic fungus of energy.To excavate more excellent growth-promoting endogenetic fungus of dendrobium candidum, on the basis of early-stage Study, this research plan from
Screening has the bacterial strain of notable growth-promoting functions to dendrobium candidum in the endogenetic fungus of Beibu Bay, guangxi mangrove separation, and combines
The morphology and molecular biological characteristics of bacterial strain carry out taxology identification to it, to be Guangxi advantage mangrove Nei Shengzhen
Utilization of the bacterium in dendrobium candidum production provide technical support.
【The content of the invention】
In view of the above-mentioned excellent growth-promoting endogenetic fungus of the more dendrobium candidums of excavation, promotes Guangxi advantage mangrove endogeny eumycete
Utilization in dendrobium candidum production, present invention aims at provide a kind of dark color have every endogenetic fungus HS40 and its
Application in dendrobium candidum production.
To reach above-mentioned purpose, the technical solution adopted in the present invention is：Dark color has every endogenetic fungus HS40, deposit number
For CGMCC NO.14345.
In the present invention, as further explanation, described dark color has the separation every endogenetic fungus HS40 to include following step
Suddenly：
(1) sample collection：Selection mangrove dominant tree-Bruguiera conjugata is sampled, using five point sampling methods gather Bruguiera conjugata root,
Stem, leaf portion tissue, are put into 4 DEG C of ice chest and save backup；
(2) configuration of culture medium：Strain isolation culture medium uses 1/2 maize powder medium；Pure medium uses malt
Juice culture medium or potato culture；Host plant-tamato seed vernalization uses pure agar medium；Rearing tomatoes with
And culture hypha,hyphae uses oat medium；
(3) fungi isolates and purifies：The sample tissue collected is cleaned and surface sterilization, after surface sterilization
Plant tissue block is placed on strain isolation culture medium flat plate, per 3 pieces of tissue blocks of ware, each ware of sample 5, in 25 DEG C of cultures day by day
See whether that mycelia grows, be transferred in time on pure medium after mycelia grows from tissue block position, obtain purifying
Fungal bacterial strain；
(4) screening of endogenetic fungus：To exclude pathogenic bacterium and saprophytic bacteria, according to colonial morphology, conidial fructification by step
(3) fungal bacterial strain of the purifying described in is grouped, and every group takes one plant of representative strain to carry out Pathogenicity, specific step at random
It is rapid as follows：
(a) inoculation, per 3 fungus blocks of ware, is cultivated 7-14d, it is standby obtains cultured bacterium colony in oat medium；
(b) epiphyte pathogenic detection is urged with host plant-tamato seed through surface sterilization, on pure agar medium
Bud, after seed is sent out into seedling on transplanting to cultured bacterium colony；
(c) transplant 1 plant of sterile tomato plants respectively on each bacterium colony, per the young plant of ware 3, culture dish is put into tissue culture bottle
In, it is 26 DEG C, co-cultured in the incubator that intensity of illumination is 1000-1200Lx and light application time is 16h/d in temperature；Each
Bacterial strain is inoculated with 12 plants of tomato plants altogether, i.e., each bacterial strain sets 4 repetitions；
(d) using the processing of non-inoculating strain, the tomato plants directly transplanted as control；
(e) incidence of tomato plants is recorded after 14d, and is put into 50 DEG C of baking ovens and dries after tomato root is cleaned
Dry weight is weighed after doing to constant weight；
(f) separated again to not showing the bacterial strain processing inapparent with contrast difference of illness, biomass, it is determined that separating again
Whether bacterial strain is former inoculating strain；
(5) endogenetic fungus root colonization is observed：The tomato plants progress root without illness in above-mentioned steps (4) is chosen to cut
Piece and the blue dyeing of cotton, observe mycelia distribution, color and the form in root tissue under the microscope, preliminary clear and definite endogenetic fungus
Colonize situation.
In the present invention, as further explanation, described dark color have every endogenetic fungus HS40 authentication method include with
Lower step：
(1) by morphological observation, its classification position is tentatively judged；
(2) it is that picking mycelia is directly used in PCR reactions, rRNA ITS1-5.8S-ITS4-28S subregions PCR amplifications
Using universal primer ITS1 (5 '-TCCGTAGGTGAACCTGCGG-3 ') and LR5 (5 '-TCCTGAGGGAAACTTCG-3 ')；
RRNA 18SrRNA sections PCR amplifications using primer NS1 (5 '-GTAGTCATATGCTTGTCTC-3 ') and NS4 (5 '-
CTTCCGTCAATTCCTTTAAG-3 '), PCR reaction systems：The μ L of 2 × EasyTaq PCR SuperMix 25,20 μm of ol/L are just
Each 1 μ L of anti-primer, the μ L of template DNA 1, use ddH2O is settled to 50 μ L.
In the present invention, it is as further explanation, the condition of the PCR reactions described in step (2)：It is anti-at 98 DEG C successively
2min is answered, 40s is reacted at 94 DEG C, reacts 1min at 50 DEG C and 4min is reacted at 68 DEG C, 30 circulate, finally 72
10min is reacted at DEG C.
In the present invention, as further explanation, dark color has is promoting dendrobium candidum plate culture every endogenetic fungus HS40
Application in seedling growth.
In the present invention, as further explanation, dark color has is promoting the life of dendrobium candidum Potted orchard every endogenetic fungus HS40
Application in length.
In the present invention, as further explanation, the dendrobium candidum seedling to being vaccinated with HS40 bacterial strains in plate culture, enter
Row root is cut into slices and the blue dyeing of cotton, mycelia distribution, color and the form in root tissue is observed under the microscope, with clear and definite HS40
Bacterial strain colonizes situation.
In the present invention, as further explanation, the blue dyeing of described cotton root tissue dyeing liquor used is by following things
Matter forms：Mass fraction is 1% cotton indigo plant 1mL, and mass fraction is 50% aqueous acetic acid 199mL.
In the present invention, liquid is enclosed by following substances group as further explanation, the blue dyeing of described cotton fungi used
Into：Phenol,Solid 20g, D-lactic acid 16mL, glycerine 31mL, sterilized water 20mL.
The invention has the advantages that：
(1) it is that having to dendrobium candidum of finding first is obvious that dark color of the present invention, which has every endogenetic fungus HS40 bacterial strains,
The Beibu Bay, guangxi mangrove endogeny eumycete of growth-promoting effect, has filled up the blank of presently relevant area research.
(2) dark color have every endogenetic fungus HS40 bacterial strains be Guangxi advantage mangrove endogeny eumycete dendrobium candidum production on
Utilization provide resource guarantee.
【Brief description of the drawings】
Fig. 1 is HS40 bacterial strain colony morphological observation figures；
Fig. 2 is HS40 bacterial strain spore structure observation figures；
Fig. 3 is based on HS40 bacterial strains and from the fungi ITS1-5.8S rRNA-ITS2-28S rRNA of GenBank phases
The NJ phylogenetic trees of gene order structure；
Fig. 4 is the NJ systems based on HS40 bacterial strains and from the fungi 18S rRNA gene orders structure of GenBank phases
Development tree；
Fig. 5 is growing way control after plate culture dendrobium candidum seedling inoculation HS40 bacterial strains；
Fig. 6 is growing way control after potted plant dendrobium candidum seedling inoculation HS40 bacterial strains；
Fig. 7 is that HS40 bacterial strains colonize situation in dendrobium candidum root.
【Embodiment】
In order to facilitate the understanding of the purposes, features and advantages of the present invention, below in conjunction with the accompanying drawings to the present invention
Embodiment be described in detail.Many details are elaborated in the following description in order to fully understand this
Invention.But the invention can be embodied in many other ways as described herein, those skilled in the art can be
Without prejudice to doing similar improvement in the case of intension of the present invention, therefore the present invention is not limited to the specific embodiments disclosed below.
Embodiment：
Embodiment 1：
Dark color has the separation every endogenetic fungus HS40
(1) mangrove sample collection
Mangrove wolf tree of certain scale is selected in Beihai, Guangxi mountain pass country's mangrove forest ecological nature reserve area
Kind-Bruguiera conjugata is sampled, and using five point sampling methods, distance about 2m between each point, collection Bruguiera conjugata root, stem, leaf portion tissue, is put
Enter 4 DEG C of ice chests temporarily to save backup.
(2) culture medium is prepared
Strain isolation culture medium uses 1/2 maize powder medium；Using malt extract medium, (malt extracts pure medium
Thing 10.0 g/L, yeast extract 2.0g/L, corn meal agar 8.5g/L, agar 7.5g/L) or potato culture (potato
200.0g/L, glucose 20.0g/L, agar 15.0g/L)；Host plant-tamato seed vernalization uses pure agar culture
Base；Rearing tomatoes and culture hypha,hyphae using oat medium (oatmeal 10.0g/L, agar 14.0g/L,
MgSO4·7H2O1.0g/L, KH2PO41.5g/L, NaNO3 1.0g/L)。
(3) fungi isolates and purifies
The sample tissue collected is cleaned and surface sterilization.The surface sterilization of plant tissue：75% ethanol vibrates
10s is sterilized, then sterilization 1-5min (wherein root 3min, stem 5min, leaf 1min) is vibrated with 1% sodium hypochlorite, uses rinsed with sterile water
It is positioned over after more than 3 times on aseptic filter paper and sucks excessive moisture, is placed on superclean bench overnight, dries；After surface sterilization
Plant tissue block be placed on 1/2CM culture medium flat plates, per ware 3 pieces of tissue blocks, each ware of sample 5, seen day by day in 25 DEG C of cultures
Whether examine has mycelia to grow, be transferred in time after mycelia grows from tissue block position malt extract medium (or potato culture
Base) on, obtain the fungal bacterial strain of purifying.
(4) screening of endogenetic fungus
To exclude pathogenic bacterium and saprophytic bacteria, step (3) separated bacterial strain is divided according to colonial morphology, conidial fructification
Group, every group takes one plant of representative strain to be used for Pathogenicity at random.By inoculation in oat medium, per 3 fungus blocks of ware,
It is standby after culture 7d.Epiphyte pathogenic detection is with host plant-tamato seed through surface sterilization, on pure agar medium
Vernalization, after seed is sent out into seedling on transplanting to cultured bacterium colony.Transplant 1 plant of sterile tomato respectively on each bacterium colony
Seedling, per ware 3 young plant, culture dish is put into tissue culture bottle, in temperature be 26 DEG C, intensity of illumination be 1000lx and light application time
To be co-cultured in 16h/d incubator；Each bacterial strain is inoculated with 12 plants of tomato plants altogether, i.e., each bacterial strain sets 4 repetitions；With
The processing of non-inoculating strain, the tomato plants directly transplanted is control.The incidence (0 of tomato plants is recorded after 14d
Level：Without disease symptom；1 grade：Aetiolation, growing way are slightly worse；2 grades：Plant substantially short and small yellow；3 grades：Plant is withered or dead
Die), and by tomato root clean after be put into be dried to constant weight in 50 DEG C of baking ovens after weigh dry weight.To not showing illness, biology
Amount bacterial strain processing inapparent with contrast difference is separated again, it is determined that whether isolated strains are former inoculating strain again.
(5) endogenetic fungus root colonization is observed
Choose the tomato plants without illness in above-mentioned (4) and carry out root section and the blue dyeing of cotton, observe under the microscope
Mycelia distribution, color and form in root tissue.Preliminary clear and definite endogenetic fungus colonizes situation.Coloring agent：1. root tissue contaminates
Color liquid：Mass fraction is 1% cotton indigo plant 1mL, and mass fraction is 50% aqueous acetic acid 199mL；2. fungi encloses liquid：Gu
Body phenol 20g, D-lactic acid 16mL, glycerine 31mL, sterilized water 20mL.
Embodiment 2：
Dark color has the separation every endogenetic fungus HS40
(1) mangrove sample collection
Mangrove wolf tree of certain scale is selected in Beihai, Guangxi mountain pass country's mangrove forest ecological nature reserve area
Kind-Bruguiera conjugata is sampled, and using five point sampling methods, distance about 2m between each point, collection Bruguiera conjugata root, stem, leaf portion tissue, is put
Enter 4 DEG C of ice chests temporarily to save backup.
(2) culture medium is prepared
Strain isolation culture medium uses 1/2 maize powder medium；Using malt extract medium, (malt extracts pure medium
Thing 10.0 g/L, yeast extract 2.0g/L, corn meal agar 8.5g/L, agar 7.5g/L) or potato culture (potato
200.0g/L, glucose 20.0g/L, agar 15.0g/L)；Host plant-tamato seed vernalization uses pure agar culture
Base；Rearing tomatoes and culture hypha,hyphae using oat medium (oatmeal 10.0g/L, agar 14.0g/L,
MgSO4·7H2O1.0g/L, KH2PO41.5g/L, NaNO3 1.0g/L)。
(3) fungi isolates and purifies
The sample tissue collected is cleaned and surface sterilization.The surface sterilization of plant tissue：75% ethanol vibrates
20s is sterilized, then sterilization 1-5min (wherein root 3min, stem 5min, leaf 1min) is vibrated with 1% sodium hypochlorite, uses rinsed with sterile water
It is positioned over after more than 3 times on aseptic filter paper and sucks excessive moisture, is placed on superclean bench overnight, dries；After surface sterilization
Plant tissue block be placed on 1/2CM culture medium flat plates, per ware 3 pieces of tissue blocks, each ware of sample 5, seen day by day in 25 DEG C of cultures
Whether examine has mycelia to grow, be transferred in time after mycelia grows from tissue block position malt extract medium (or potato culture
Base) on, obtain the fungal bacterial strain of purifying.
(4) screening of endogenetic fungus
To exclude pathogenic bacterium and saprophytic bacteria, step (3) separated bacterial strain is divided according to colonial morphology, conidial fructification
Group, every group takes one plant of representative strain to be used for Pathogenicity at random.By inoculation in oat medium, per 3 fungus blocks of ware,
It is standby after culture 10d.Epiphyte pathogenic detection is with host plant-tamato seed through surface sterilization, on pure agar medium
Vernalization, after seed is sent out into seedling on transplanting to cultured bacterium colony.Transplant 1 plant of sterile tomato respectively on each bacterium colony
Seedling, per ware 3 young plant, culture dish is put into tissue culture bottle, in temperature be 26 DEG C, intensity of illumination be 1100lx and light application time
To be co-cultured in 16h/d incubator；Each bacterial strain is inoculated with 12 plants of tomato plants altogether, i.e., each bacterial strain sets 4 repetitions；With
The processing of non-inoculating strain, the tomato plants directly transplanted is control.The incidence (0 of tomato plants is recorded after 14d
Level：Without disease symptom；1 grade：Aetiolation, growing way are slightly worse；2 grades：Plant substantially short and small yellow；3 grades：Plant is withered or dead
Die), and by tomato root clean after be put into be dried to constant weight in 50 DEG C of baking ovens after weigh dry weight.To not showing illness, biology
Amount bacterial strain processing inapparent with contrast difference is separated again, it is determined that whether isolated strains are former inoculating strain again.
(5) endogenetic fungus root colonization is observed
Choose the tomato plants without illness in above-mentioned (4) and carry out root section and the blue dyeing of cotton, observe under the microscope
Mycelia distribution, color and form in root tissue.Preliminary clear and definite endogenetic fungus colonizes situation.Coloring agent：1. root tissue contaminates
Color liquid：Mass fraction is 1% cotton indigo plant 1mL, and mass fraction is 50% aqueous acetic acid 199mL；2. fungi encloses liquid：Gu
Body phenol 20g, D-lactic acid 16mL, glycerine 31mL, sterilized water 20mL.
Embodiment 3：
Dark color has the separation every endogenetic fungus HS40
(1) mangrove sample collection
Mangrove wolf tree of certain scale is selected in Beihai, Guangxi mountain pass country's mangrove forest ecological nature reserve area
Kind-Bruguiera conjugata is sampled, and using five point sampling methods, distance about 2m between each point, collection Bruguiera conjugata root, stem, leaf portion tissue, is put
Enter 4 DEG C of ice chests temporarily to save backup.
(2) culture medium is prepared
Strain isolation culture medium uses 1/2 maize powder medium；Using malt extract medium, (malt extracts pure medium
Thing 10.0 g/L, yeast extract 2.0g/L, corn meal agar 8.5g/L, agar 7.5g/L) or potato culture (potato
200.0g/L, glucose 20.0g/L, agar 15.0g/L)；Host plant-tamato seed vernalization uses pure agar culture
Base；Rearing tomatoes and culture hypha,hyphae using oat medium (oatmeal 10.0g/L, agar 14.0g/L,
MgSO4·7H2O1.0g/L, KH2PO41.5g/L, NaNO3 1.0g/L)。
(3) fungi isolates and purifies
The sample tissue collected is cleaned and surface sterilization.The surface sterilization of plant tissue：75% ethanol vibrates
30s is sterilized, then sterilization 1-5min (wherein root 3min, stem 5min, leaf 1min) is vibrated with 1% sodium hypochlorite, uses rinsed with sterile water
It is positioned over after more than 3 times on aseptic filter paper and sucks excessive moisture, is placed on superclean bench overnight, dries；After surface sterilization
Plant tissue block be placed on 1/2CM culture medium flat plates, per ware 3 pieces of tissue blocks, each ware of sample 5, seen day by day in 25 DEG C of cultures
Whether examine has mycelia to grow, be transferred in time after mycelia grows from tissue block position malt extract medium (or potato culture
Base) on, obtain the fungal bacterial strain of purifying.
(4) screening of endogenetic fungus
To exclude pathogenic bacterium and saprophytic bacteria, step (3) separated bacterial strain is divided according to colonial morphology, conidial fructification
Group, every group takes one plant of representative strain to be used for Pathogenicity at random.By inoculation in oat medium, per 3 fungus blocks of ware,
It is standby after culture 14d.Epiphyte pathogenic detection is with host plant-tamato seed through surface sterilization, on pure agar medium
Vernalization, after seed is sent out into seedling on transplanting to cultured bacterium colony.Transplant 1 plant of sterile tomato respectively on each bacterium colony
Seedling, per ware 3 young plant, culture dish is put into tissue culture bottle, in temperature be 26 DEG C, intensity of illumination be 1200lx and light application time
To be co-cultured in 16h/d incubator；Each bacterial strain is inoculated with 12 plants of tomato plants altogether, i.e., each bacterial strain sets 4 repetitions；With
The processing of non-inoculating strain, the tomato plants directly transplanted is control.The incidence (0 of tomato plants is recorded after 14d
Level：Without disease symptom；1 grade：Aetiolation, growing way are slightly worse；2 grades：Plant substantially short and small yellow；3 grades：Plant is withered or dead
Die), and by tomato root clean after be put into be dried to constant weight in 50 DEG C of baking ovens after weigh dry weight.To not showing illness, biology
Amount bacterial strain processing inapparent with contrast difference is separated again, it is determined that whether isolated strains are former inoculating strain again.
(5) endogenetic fungus root colonization is observed
Choose the tomato plants without illness in above-mentioned (4) and carry out root section and the blue dyeing of cotton, observe under the microscope
Mycelia distribution, color and form in root tissue.Preliminary clear and definite endogenetic fungus colonizes situation.Coloring agent：1. root tissue contaminates
Color liquid：Mass fraction is 1% cotton indigo plant 1mL, and mass fraction is 50% aqueous acetic acid 199mL；2. fungi encloses liquid：Gu
Body phenol 20g, D-lactic acid 16mL, glycerine 31mL, sterilized water 20mL.
Embodiment 4：
Dark color has the identification every endogenetic fungus HS40
(1) morphological observation of bacterial strain
1. will be inoculated in for examination fungal bacterial strain on malt extract medium, culture observes colony morphology characteristic after 3 weeks；2. use nothing
Cap slide oblique cutting enters on oat medium, is inoculated with culture medium side for examination fungal bacterial strain, and 25 DEG C are cultivated 2-4 weeks, are treated obvious
When seeing that mycelia life on the cover slip, slide is taken out, carries out observing mycelia and spore growth feelings under light microscope
Condition.
HS40 bacterial strain colony morphological observation results are shown in Fig. 1, and HS40 bacterial strain spore structure morphologic observation results are shown in Fig. 2.
As shown in Figure 1：Bacterial strain long speed on malt extract medium is general, colony diameter 25mm or so after 25 DEG C of cultures 2 weeks,
Yellowish-brown is circular to brown, carpet shape.
As shown in Figure 2：Mycelia fawn, have every 1.4-4.0 μm of diameter；Conidiophore yellowish-brown, more points of 2-
Every uprightly, branch, size are not 23.2-154.2 × 2.2-5.5 μm；Conidium fawn to yellowish-brown, strip,
Long club-like or oblong, 0-4 separation, a diameter of 8.3-60.2 × 1.8-3.6 μm.Features described above and Zasmidium's
Morphological feature is consistent substantially.
(2) molecular biology identification of bacterial strain
Mycelia on picking CMMY plates is directly used in PCR reactions, rRNA ITS1-5.8S-ITS4-28S subregions
PCR amplifications using universal primer ITS1 (5 '-TCCG TAGGTGAACCTGCGG-3 ') and LR5 (5 '-
TCCTGAGGGAAACTTCG-3′)；RRNA 18S rRNA sections PCR amplification using primer NS1 (5 '-
GTAGTCATATGCTTGTCTC-3 ') and NS4 (5 '-CTTC CGTC AATT CCTT TAAG-3 '), PCR reaction systems：2×
EasyTaq PCR SuperMix 25 μ L, 20 μm of each 1 μ L of the positive anti-primers of ol/L, the μ L of template DNA 1 (a small amount of mycelia of picking), use
ddH2O is settled to 50 μ L.PCR reaction conditions：2min is reacted at 98 DEG C successively, reacts 40s at 94 DEG C, it is anti-at 50 DEG C
1min is answered, 4min, 30 circulations are reacted at 68 DEG C；Finally 10min is reacted at 72 DEG C.PCR primer is directly surveyed after purification
Sequence, examining order are completed by Hua Da Gene science Services Co., Ltd.
The sequence that experiment obtains carries out homologous comparison in GenBank, downloads most like sequence, uses Clustalx
The softwares of 1.81 and MEGA 6.0 are based on Kimura two-parameter models structure Neighbor-Joining phylogenetic trees, warp
The reliability of 1000 loop checking system trees of Bootstrap, the results are shown in Table 1.
Based on HS40 bacterial strains and from the fungi ITS1-5.8S rRNA-ITS2-28S rRNA gene sequences of GenBank phases
Arrange structure NJ phylogenetic trees with based on HS40 bacterial strains and from the fungi 18S rRNA gene order structures of GenBank phases
The NJ phylogenetic trees built are as shown in Figure 3 and Figure 4.
Table 1：
As shown in Figure 3 and Table 1：Based on HS40 bacterial strains and similar fungi ITS 1-5.8S rRNA-ITS2-28S rRNA bases
Because the NJ phylogenetic trees of sequence construct are shown, HS40 is with 99% supporting rate and 98% sequence similarity and Zasmidium
Citri gathers for one kind；
As shown in Figure 4 and Table 1：NJ phylogenetic trees based on 18S rRNA gene orders structure show that HS40 is with 99%
Supporting rate and 99% sequence similarity and Zasmidium citri griseum gather for one kind.
Combining form and molecular biology identification result, are set to spherical cavity Cordycepps (Mycosphaerellaceae) by HS40
Zasmidium sp..
Embodiment 5：
Plate is tested：
Oat medium (oatmeal 10g/L, agar 18g/L, MgSO are inoculated in after bacterial strain HS40 is activated4·7H2O 1g/
L, KH2PO41.5g/L, NaNO3On 1g/L), 3 fungus blocks are inoculated with per ware, it is standby after bacterium colony is grown up.Select plant height and healthy and strong
On basically identical sterile dendrobium candidum seedling replanting to the bacterium colony of degree, 1 plant of sterile iron sheet stone is transplanted respectively on each bacterium colony
Dry measure used in former times seedling, per 3 plants of ware, culture dish is put into tissue culture bottle, co-cultured in incubator, cultivation temperature is 26 DEG C, intensity of illumination
It is 16h/d for 1000-1200lx and light application time, the growth indexes such as plant height is measured after 60d.Using non-inoculating strain as control at
Reason.Each 4 glasss of processing, if 4 repetitions.
Plate cultivation results are shown in Table 2, after plate culture dendrobium candidum seedling inoculation HS40 bacterial strains growing way control see Fig. 5.
Table 2：
| Plant height (mm) | Root long (cm) | Stem diameter (mm) | Fresh weight (mg) | Dry weight (mg) | |
| HS40 | 56.67A | 2.77 | 4.21 | 790.1 | 125.4A |
| CK | 46.77B | 2.50 | 3.28 | 735.0 | 52.1C |
Remarks：1. different capitalizations represent difference up to the 1% pole level of signifiance respectively with after column data；2. upper table is only listed
HS40 and the results of analysis of variance of control (totally 5 bacterial strains and 1 control).
As shown in table 2：It is inoculated with dendrobium candidum seedling plant height, stem diameter, plant fresh weight and the dry weight relatively control increasing respectively of the bacterial strain
Add 21.2%, 28.4%, 7.5% and 140.7%, wherein, plant height and dry weight reach the pole level of signifiance, explanation with contrast difference
The growth of dendrobium candidum seedling has obvious facilitation during endogenetic fungus HS40 tests for plate, also further illustrates
Endogenetic fungus HS40 bacterial strains are the endogenetic fungus for having to dendrobium candidum obvious growth-promoting effect.
Embodiment 6：
Pot experiment：
(1) tissue-cultured seedling plantation, maintenance：Candidum tissue culturing seedling for the seed selection of Guangxi Academy of Agricultural Sciences flowers institute quality 784.
Candidum tissue culturing seedling is planted with pasture and water, 5 plants/glass, plants 320 glasss altogether；
(2) fungi-releasing methods：Dendrobium candidum seed starts to pour bacterium when planting 30d.20 processing are set (including i.e. comprising HS40
19 bacterial strains and control CK), each 4 glasss of processing, 4 repetitions (it is a repetition that selection, which grows more consistent cup seedling, as far as possible), one
Individual 16 glasss of processing.If 4 repetitions.Fungi-releasing methods：20ml/ cups are poured, interval 15d is poured once, poured altogether 3 times again.Separately not pour bacterium solution
For control treatment；
(3) growth indexes determine：180d is sampled after pouring bacterium for the first time, and investigation different disposal dendrobium candidum Potted orchard survives
Rate, record plant height, stem diameter, root long simultaneously determine the relative growths such as fresh weight, dry weight, leaf growth element IAA, stem polyoses content and referred to
Mark；
(4) statistical analysis：Analyzed using Excel 2003 and the softwares of DPS 6.55, test data is with average value ± mark
Standard represents by mistake.
Results from pot experiment test is shown in Table 3, after potted plant dendrobium candidum seedling inoculation HS40 bacterial strains growing way control see Fig. 6, HS40 bacterial strains
See Fig. 7 in dendrobium candidum root colonization situation.
Table 3：
Remarks：1. do not represent difference up to the 1% pole level of signifiance and 5% notable water with the mother stock that writes of different size after column data
It is flat.2. upper figure only lists the results of analysis of variance (totally 20 bacterial strains and 1 control) of HS40 and control.
As shown in table 3, the dendrobium candidum Potted orchard tiller number of bacterial strain processing, plant fresh weight, plant weights and stem are more
Sugared content relatively control increase by 400.0%, 94.8%, 56.8%, 15.8% respectively, wherein tiller number reach notable with contrast difference
Level, fresh weight, dry weight and stem polyoses content then respectively reach the pole level of signifiance, illustrate that HS40 can not only promote iron sheet stone
The growth of dry measure used in former times Potted orchard, dendrobium candidum active ingredient (polysaccharide) content can also be improved, and then improve its quality.Determine indoles
Acetic acid (IAA) content finds, it is higher by 16.2% than compareing to be inoculated with the dendrobium candidum blade of the bacterial strain IAA contents, and difference is up to extremely showing
Level is write, illustrates that HS40 can promote dendrobium candidum blade IAA synthesis, and then promote Growth of Dendrobium candidum and development；Also one is entered
Step illustrates that endogenetic fungus HS40 bacterial strains are the endogenetic fungus for having to dendrobium candidum obvious growth-promoting effect.
Described above is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, the equal change completed or modification change under the technical spirit suggested by all present invention, all should belong to
Cover the scope of the claims in the present invention.
Sequence table
<110>Institute of Microbiology, Guangxi Academy of Agricultural Sciences
<120>Dark color has every endogenetic fungus HS40 and its application on dendrobium candidum
<130> ZP17101962/WSW
<141> 2017-08-30
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 19
<212> DNA
<213>Artificial sequence (zasmidium sp.)
<400> 1
tccgtaggtg aacctgcgg 19
<210> 2
<211> 17
<212> DNA
<213>Artificial sequence (zasmidium sp.)
<400> 2
tcctgaggga aacttcg 17
<210> 3
<211> 19
<212> DNA
<213>Artificial sequence (zasmidium sp.)
<400> 3
gtagtcatat gcttgtctc 19
<210> 4
<211> 20
<212> DNA
<213>Artificial sequence (zasmidium sp.)
<400> 4
cttccgtcaa ttcctttaag 20
Claims (9)
1. dark color has every endogenetic fungus HS40, it is characterised in that：Deposit number is CGMCC NO.14345.
2. dark color has every endogenetic fungus HS40, it is characterised in that：Described dark color have every endogenetic fungus HS40 separation include with
Lower step：
(1) sample collection：Selection mangrove dominant tree-Bruguiera conjugata is sampled, using five point sampling methods gather Bruguiera conjugata root, stem,
Leaf portion tissue, it is put into 4 DEG C of ice chest and saves backup；
(2) configuration of culture medium：Strain isolation culture medium uses 1/2 maize powder medium；Pure medium is trained using brewer's wort
Support base or potato culture；Host plant-tamato seed vernalization uses pure agar medium；Rearing tomatoes and culture
Hypha,hyphae uses oat medium；
(3) fungi isolates and purifies：The sample tissue collected is cleaned and surface sterilization, by the plant after surface sterilization
Tissue block is placed on strain isolation culture medium flat plate, and per 3 pieces of tissue blocks of ware, each ware of sample 5, in 25 DEG C of cultures, observation is day by day
It is no to there is mycelia to grow, it is transferred in time on pure medium after mycelia grows from tissue block position, obtains the fungi bacterium of purifying
Strain；
(4) screening of endogenetic fungus：To exclude pathogenic bacterium and saprophytic bacteria, according to colonial morphology, conidial fructification by step (3) institute
The fungal bacterial strain for the purifying stated is grouped, and every group takes one plant of representative strain to carry out Pathogenicity at random, comprises the following steps that：
(a) inoculation, per 3 fungus blocks of ware, is cultivated 7-14d, it is standby obtains cultured bacterium colony in oat medium；
(b) epiphyte pathogenic detection is treated with host plant-tamato seed through surface sterilization, the vernalization on pure agar medium
After seed is sent out into seedling on transplanting to cultured bacterium colony；
(c) transplant 1 plant of sterile tomato plants respectively on each bacterium colony, per the young plant of ware 3, culture dish be put into tissue culture bottle,
It is 26 DEG C, co-cultured in the incubator that intensity of illumination is 1000-1200Lx and light application time is 16h/d in temperature；Each bacterial strain
12 plants of tomato plants are inoculated with altogether, i.e., each bacterial strain sets 4 repetitions；
(d) using the processing of non-inoculating strain, the tomato plants directly transplanted as control；
(e) incidence of tomato plants is recorded after 14d, and is put into 50 DEG C of baking ovens and is dried to after tomato root is cleaned
Dry weight is weighed after constant weight；
(f) separated again to not showing the bacterial strain processing inapparent with contrast difference of illness, biomass, it is determined that isolated strains again
Whether it is former inoculating strain；
(5) endogenetic fungus root colonization is observed：Choose the tomato plants without illness in above-mentioned steps (4) carry out root section and
The blue dyeing of cotton, observes mycelia distribution, color and form in root tissue under the microscope, and preliminary clear and definite endogenetic fungus colonizes feelings
Condition.
3. dark color has every endogenetic fungus HS40, it is characterised in that：Described dark color has the authentication method bag every endogenetic fungus HS40
Include following steps：
(1) by morphological observation, its classification position is tentatively judged；
(2) it is that picking mycelia is directly used in PCR reactions, rRNA ITS1-5.8S-ITS4-28S subregions PCR amplifications use
Universal primer ITS1 (5 '-TCCGTAGGTGAACCTGCGG-3 ') and LR5 (5 '-TCCTGAGGGAAACTTCG-3 ')；RRNA's
18SrRNA sections PCR amplifications using primer NS1 (5 '-GTAGTCATATGCTTGTCTC-3 ') and NS4 (5 '-
CTTCCGTCAATTCCTTTAAG-3 '), PCR reaction systems：The μ L of 2 × EasyTaq PCR SuperMix 25,20 μm of ol/L are just
Each 1 μ L of anti-primer, the μ L of template DNA 1, use ddH2O is settled to 50 μ L.
4. dark color according to claim 3 has every endogenetic fungus HS40, it is characterised in that：PCR reactions described in step (2)
Condition be：2min is reacted at 98 DEG C successively, 40s is reacted at 94 DEG C, reacts 1min at 50 DEG C and reacted at 68 DEG C
4min, 30 circulations, finally reacts 10min at 72 DEG C.
5. the application every endogenetic fungus HS40 in dendrobium candidum production is had according to any described dark colors of claim 1-4, its
It is characterised by：Dark color has the application on the growth of dendrobium candidum plate cultivating seedling is promoted every endogenetic fungus HS40.
6. the application every endogenetic fungus HS40 in dendrobium candidum production is had according to any described dark colors of claim 1-4, its
It is characterised by：Dark color has the application on the growth of dendrobium candidum Potted orchard is promoted every endogenetic fungus HS40.
7. having the application every endogenetic fungus HS40 in dendrobium candidum production according to any described dark color of claim 5, it is special
Sign is：Dendrobium candidum seedling to being vaccinated with HS40 bacterial strains in plate culture, root section and the blue dyeing of cotton are carried out, micro-
Mycelia distribution, color and form in Microscopic observation root tissue, situation is colonized with clear and definite HS40 bacterial strains.
8. having the application every endogenetic fungus HS40 in dendrobium candidum production according to any described dark color of claim 7, it is special
Sign is：The blue dyeing of described cotton root tissue dyeing liquor used is made up of following substances：The cotton that mass fraction is 1% is blue
1mL, mass fraction are 50% aqueous acetic acid 199mL.
9. having the application every endogenetic fungus HS40 in dendrobium candidum production according to any described dark color of claim 7, it is special
Sign is：The blue dyeing of described cotton fungi used encloses liquid and is made up of following substances：Phenol,Solid 20g, D-lactic acid 16mL,
Glycerine 31mL, sterilized water 20mL.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108719336A (en) * | 2018-05-18 | 2018-11-02 | 中国林业科学研究院林业研究所 | Carry on a shoulder pole the method that Pseudomonas fungi promotes orchid growth in angle |
| CN110205249A (en) * | 2019-06-12 | 2019-09-06 | 中国矿业大学(北京) | A kind of method promoting plant growth and its rod method fungi used |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101822211A (en) * | 2009-09-03 | 2010-09-08 | 昆明优利丰园科技开发有限公司 | Tissue cultivating method of dendrobium officinale growing in Yunnan |
| CN103981102A (en) * | 2014-05-23 | 2014-08-13 | 广西壮族自治区农业科学院微生物研究所 | DSE (Dark septate endophyte) bacterial strain 24L-4 and application thereof in dendrobium officinale production |
| US20160143236A1 (en) * | 2013-07-08 | 2016-05-26 | South China Botanical Garden, Chinese Academy Of Sciences | Dendrobium in vitro crossbreeding method |
| CN105918127A (en) * | 2016-05-20 | 2016-09-07 | 武汉理工大学 | Method for building DSE-plant symbiosis system based on DSE rapid expanding propagation |
| CN107083335A (en) * | 2017-04-25 | 2017-08-22 | 鲁东大学 | One plant of DSE fungi and the method for the quick Mycorrhizal of blueberry tissue culture seedling |
-
2017
- 2017-09-15 CN CN201710833508.3A patent/CN107460133B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101822211A (en) * | 2009-09-03 | 2010-09-08 | 昆明优利丰园科技开发有限公司 | Tissue cultivating method of dendrobium officinale growing in Yunnan |
| US20160143236A1 (en) * | 2013-07-08 | 2016-05-26 | South China Botanical Garden, Chinese Academy Of Sciences | Dendrobium in vitro crossbreeding method |
| CN103981102A (en) * | 2014-05-23 | 2014-08-13 | 广西壮族自治区农业科学院微生物研究所 | DSE (Dark septate endophyte) bacterial strain 24L-4 and application thereof in dendrobium officinale production |
| CN105918127A (en) * | 2016-05-20 | 2016-09-07 | 武汉理工大学 | Method for building DSE-plant symbiosis system based on DSE rapid expanding propagation |
| CN107083335A (en) * | 2017-04-25 | 2017-08-22 | 鲁东大学 | One plant of DSE fungi and the method for the quick Mycorrhizal of blueberry tissue culture seedling |
Non-Patent Citations (5)
| Title |
|---|
| D.G. KNAPP等: "Dark septate endophytic pleosporalean genera from semiarid areas", 《PERSOONIA》 * |
| XIE, L.: "ACCESSION：KY576814.1，Fungal sp. HS40 LX-2016 small subunit ribosomal RNA gene, partial sequence", 《GENBANK》 * |
| XIE, L.: "KY496825.1，Fungal sp. HS40 LX-2016 small subunit ribosomal RNA gene, partial sequence; internal transcribed spacer 1, 5.88 ribosomal RNA gene, and internal transcribed spacer 2, complete sequence; and large subunit ribosomal RNA gene, partial sequence", 《GENBANK》 * |
| 刘茂军等: "深色有隔内生真菌（DSE）研究进展", 《菌物学报》 * |
| 张艳等: "广西北部湾红树植物内生真菌多样性", 《微生物学通报》 * |
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| CN111808758A (en) * | 2019-08-30 | 2020-10-23 | 广西壮族自治区农业科学院微生物研究所 | Crop growth-promoting endophytic fungus and application thereof |
| CN111808758B (en) * | 2019-08-30 | 2021-03-05 | 广西壮族自治区农业科学院微生物研究所 | Crop growth-promoting endophytic fungus and application thereof |
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